This application is the National Stage filed under 35 USC 371 of PCT/EP98/05025, filed Aug. 7, 1998.
The invention relates to substituted tetrahydropyran derivatives, processes for their preparation, their use as a pharmaceutical or diagnostic and pharmaceutical comprising them.
Peptides and peptide mimetics are a valuable aid for the discovery of new lead structures and the identification of potential active compounds. By fixing side chains in a rigid structure (scaffold), it is hoped, in comparison with the conformationally more flexible peptide chain, for an increase in the affinity of this conformationally fixed ligand for the receptor.
Very different structural units are already finding use as peptide mimetics.
Owing to their polyvalency and their defined spatial arrangement, carbohydrate units should be particularly highly suitable as structural units for peptide mimetics.
Thus, it has recently been shown that a specific monosaccharide mimics, as a conformationally fixed structure, the spatial arrangement of a certain cyclopeptide, somatostatin (K. C. Nicolaou, J. I. Trujillo, K. Chibale, Tetrahedron 1997, 53, 8751-8778).
In this connection, starting from a glucose derivative having standard protective groups, a restricted variation of the simply accessible anomeric hydroxyl function and the C-6 hydroxyl function was carried out. The synthesis strategy described there starts from already known sugar units and is restricted by the protective group:strategy to a narrow application range of somatostatin. At the same time, the method is not transferable to the targeted variation of the structural unit by solid-phase synthesis.
The previous syntheses of carbohydrate derivatives in solution or in the form of substance libraries on a solid phase concentrated, in particular, on the synthesis of oligosaccharides or glycopeptides (L. DeNapoli et al., Tetrahedron Letters 1996, 37, 5007-5010; S. J. Danishefsky et al., Science 1995, 269, 202-204, J. J. Krepinski et al., J. Am. Chem. Soc., 1991, 113, 5095-5097).
Compounds synthesized from oligosaccharide or glycopeptide units are, however, of only very restricted use for the discovery of lead structures or as potential active compounds on account of their complexity.
Restriction to a monosaccharide as a structural unit, however, combines the positive property of the defined spatial arrangement of the ligands with a low complexity, low molecular weight, low toxicity and further properties which are of importance for potential active compounds.
On account of the polyvalency of the monosaccharides, targeted synthesis of selectively functionalized monosaccharidesxe2x80x94both in solution and on the solid phasexe2x80x94causes great difficulties.
Variously protected carbohydrate units are likewise known as a result of the various studies on carbohydrate chemistry (see R. R. Schmidt, Pure and Appl. Chem. 1989, 61, 1257). In the intermediates described there, the hydroxyl groups are temporarily blocked more or less selectively by protective groups which are then deprotected for linkage with other protective groups, as a result of which the synthesis of di- or oligosaccharides takes place.
These intermediates or the polysugars synthesized therefrom are, however, of only restricted use for specific lead structure discovery and as potential active compounds. In some cases, these structures are relatively labile and thus not resistant to degradation or cleavage.
The linkers and activation strategies developed for the preparation of the abovementioned polysaccharides or glycopeptides (D. Kahne et al., J. Am. Chem. Soc. 1994, 116, 6953-6954) are also not generally transferable to the preparation of selectively polysubstituted monosaccharide compounds.
The specific synthesis of selectively functionalized monosaccharide derivatives therefore requires the development of a novel, completely orthogonal protective group strategy, which makes it possible to selectively remove the protective groups of all functional groups, the conditions used for this being stable to the conditions of the synthesis sequence. At the same time, these protective groups must guarantee compatibility with all reaction conditions which are necessary for synthesis in solid-phase synthesis. For synthesis on a solid phase, it is furthermore necessary to have available a linker system for linking the monosaccharide unit, preferably via the anomeric center, which is compatible with all reaction conditions and can be selectively activated. Such a strategy makes possible the specific different variation of all functionalities of the monosaccharide unit to give stable final products.
The invention thus relates to compounds of the formula I 
in which:
R1, R2, R3, R4, R5 independently of one another are
1. hydrogen;
2. (C1-C12)-alkyl;
3. (C2-C8)-alkenyl;
4. (C2-C8)-alkynyl;
5. (C1-C6)-alkylene-(C3-C10)-cycloalkyl;
6. (C0-C6)-alkylene-(C6-C12)-aryl; preferably phenyl or benzyl;
7. (C1-C6)-alkoxy;
8. (C0-C6)-alkylene-COxe2x80x94R8;
9. (C1-C6)-alkylene-(C1-C9)-heteroaryl;
10. carbamoyl;
11. xe2x80x94C(O)NR6R7;
12. xe2x80x94C(O)OR6;
13. a radical defined as in 2.-12., which is mono-, di- or polysubstituted in the alkyl moiety and/or aryl or heteroaryl moiety by a radical from the group consisting of (C1-C6)-alkyl, NO2, CN, halogen, CF3 or (C1-C6)-alkoxy;
14. a radical defined as in 6. and 9., which is substituted in the aryl or heteroaryl moiety by one, two or more halogen atoms;
R6 and R7 independently of one another are:
1. hydrogen;
2. (C1-C12)-alkyl;
3. (C2-C8)-alkenyl;
4. (C2-C8)-alkynyl;
5. (C1-C6)-alkylene-(C3-C10)-cycloalkyl;
6. (C1-C6)-alkylene-(C6-C12)-aryl; preferably benzyl;
7. (C2-C6)-alkyloxy;
8. (C0-C6)-alkylene-COxe2x80x94R8;
9. (C1-C6)-alkylene-(C1-C9)-heteroaryl;
10. (C0-C6)-alkylene-(C1-C6)-alkoxy;
11. (C3-C10)-cycloalkyl;
12. (C6-C12)-aryl, preferably phenyl;
R8 is hydrogen, (C1-C6)-alkyl, (C6-C12)-aryl or OR12;
R12 is hydrogen, (C1-C6)-alkyl or (C6-C12)-aryl; or
R2 and R3 together or R3 and R4 together or R4 and R5 together are
(C1-C3)-alkylene which can be substituted by 1 or 2 (C1-C3)-alkyl radicals or optionally substituted (C6-C12)-aryl radicals;
X is N or O;
with the proviso that R2 is not xe2x80x94C(O)OR6 when X is O;
and their physiologically tolerable salts.
Preferred compounds of the formula I are those in which the radicals R1, R2, R3, R4 and R5 do not each have the same meaning, and their physiologically tolerable salts.
Preferred compounds of the formula I are furthermore those in which only three of the radicals R1, R2, R3, R4, R5 have the same meaning, and their physiologically tolerable salts.
Particularly preferred compounds of the formula I are those in which only two of the radicals R1, R2, R3, R4, R5 have the same meaning, and their physiologically tolerable salts.
Very particularly preferred compounds of the formula I are those in which all radicals R1, R2, R3, R4, R5 have a different meaning, and their physiologically tolerable salts.
Preferred compounds of the formula I are those in which at least one of the radicals R1, R2, R3, R4, R5 is hydrogen, xe2x80x94C(O)NR6R7, (C1-C8)-alkyl, (C0-C6)-alkyl-(C6-C12)-aryl, preferably phenyl or benzyl; the aryl moiety of the (C1-C6)-alkyl-(C6-C12)-aryl radical being unsubstituted or mono-, di- or trisubstituted by (C1-C6)-alkyl, cyano, nitro, CF3, Cl, Br or (C1-C4)alkoxy, preferably methoxy, and R6 and R7 independently of one another are hydrogen, (C1-C4)-alkyl, benzyl, (C1-C3)-alkylene-(C3-C7)-cycloalkyl, (C1-C3)-alkylene-COxe2x80x94OR12, (C1-C3)-alkylene-(C1-C3)-alkoxy, phenyl, optionally substituted by one or two radicals from the group consisting of CF3, Cl, Br, F, nitro, cyano; and R12 is as defined above;
or R3 and R4 together or R4 and R5 together are xe2x80x94CH2xe2x80x94 which is substituted by 1 or 2 methyl radicals or optionally substituted phenyl radicals, and the other radicals are as defined above,
and their physiologically tolerable salts.
Preferred compounds of the formula I are also those in which X is equal to xe2x80x94Oxe2x80x94 and the other radicals are as defined above, and their physiologically tolerable salts.
The invention furthermore relates to compounds of the formula II 
in which:
R1 is a linker group which can be linked via a covalent bond to a carrier functionalized by a heteroatom, for example N, O or Cl:
R2, R3, R4, R5 independently of one another are a protective group customary in sugar chemistry;
Y is O or S, preferably S;
X is O or N, preferably O.
Protective groups customary in sugar chemistry are, for example, those such as are described, for example, in T. W. Greene, P. G. Wuts xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d, 2nd Edition, Wiley/New York, 1991.
Suitable protective groups for compounds of the formula II are, for example, silyl protective groups, e.g. TBDPS; alkoxyalkyl groups, e.g. ethoxyethyl; allyl groups; acyl groups such as acetyl or benzoyl; acetals and ketals such as isopropylidene or optionally substituted benzylidene.
Preferred compounds of the formula II are those in which the radicals R2, R3, R4 and R5 are not all the same protective group.
Preferred compounds of the formula II are furthermore those in which only two of the radicals R2, R3, R4, R5 are an identical protective group.
Very particularly preferred compounds of the formula II are those in which the radicals R2, R3, R4, R5 are each a different protective group.
Preferred compounds of the formula II are also those which have an orthogonal protective group pattern with protective groups from the following different classes:
base-labile protective groups, such as the acetate or benzoyl group;
acid-labile protective groups, such as acetal- or ketal-like protective groups such as the ethoxyethyl group;
fluoride-labile protective groups, such as the tert-butyidimethylsilyl or tert-butyidiphenylsilyl group;
a protective group removable by transition metal catalysis, such as the allyl group;
sulfur-containing protective groups, such as in the linker system.
The invention also relates to compounds of the formula II, in which:
R5 is a linker group which can be linked via a covalent bond to a carrier functionalized by a heteroatom, for example N, O or Cl;
R1, R2, R3, R4 independently of one another are a protective group customary in sugar chemistry;
Y is S or O, preferably S;
X is O or N, preferably O.
The invention also relates to compounds of the formulae IIa, IIb and IIc 
in which:
Y is S or O, preferably S;
X is O or N, preferably O;
R1 is a linker group which can be linked via a covalent bond to a carrier functionalized by a heteroatom, for example N, O or Cl;
R2 in the case in which X is equal to 0, is a base-labile protective group such as, for example, acetyl or benzoyl;
in the case in which X is equal to N, is a base-labile protective group such as, for example, a phthaloyl protective group, or DDE (1-(4,4-dimethyl-2,6-dioxocyclohexylidene-ethyl) or NDE (2-acetyl4-nitroindan-1,3-dione);
R3 is an allyl protective group;
R4 is an acid-labile protective group, such as acetal- or ketal-like protective groups, for example ethoxyethyl or SEM (2-(trimethylsilyl)ethoxymethyl);
R5 is a suitable silyl protective group, such as, for example, tert-butyldimethylsilyl or tert-butyldiphenylsilyl.
In general, suitable silyl protective groups for R5 are fluoride-labile protective groups, which are more stable, i.e. more difficult to remove, than a trimethylsilyl radical.
The invention also relates to compounds of the formula IIa, in which R4 and R5 together are an acetal- or ketal-like protective group such as, for example, isopropylidene or benzylidene and the other radicals X, Y, R1, R2 and R3 are as defined above.
The invention additionally relates to compounds of the formula IIb, in which R3 and R4 together are an acetal- or ketal-like protective group such as, for example, isopropylidene or benzylidene and the other radicals X, Y, R1, R2 and R5 are as defined above.
A suitable linker group R1 or R5 is, for example, a group of the formula III
(C1-C6)-alkylene-[Nxe2x80x94C(O)]nxe2x80x94[(C6-C12)-arylene]pxe2x80x94(C0-C6)-alkylene-C(O)R9xe2x80x83xe2x80x83(III)
in which n and p are 0 or 1, where p and n cannot simultaneously be 1;
R9 is OR10 or NR11R11, where
R10 is H, (C1-C6)-alkyl, (C1-C6)-alkyl-(C6-C12)-aryl, and
R11 independently of one another is H, (C1-C6)-alkyl, (C1-C6)-alkyl-(C6-C12)-aryl or a polymeric solid support.
Preferred compounds of the formula I or II are also those in which R2 and R3 together, or R3 and R4 together or R4 and R5 together form a benzylidene radical or isopropylidene radical and the other radicals are as defined above.
Preferred compounds of the formula I and formula II are furthermore those in which the monosaccharide structure is a glucose unit, a galactose unit or a mannose unit.
The compounds of the formula II and of the formula IIa, IIb or IIc are valuable intermediates for the preparation of compounds of the formula I.
(C1-C6)-Aryl is understood, for example, as meaning phenyl, naphthyl or biphenyl.
Alkyl, alkenyl, alkynyl, alkylene and radicals derived therefrom such as, for example, alkoxy can be straight-chain or branched, those branched radicals being preferred in which the branching site is not directly situated on the linkage site to the monosaccharide structure.
Halogen is preferably fluorine, chlorine or bromine.
A heteroaryl radical within the meaning of the present invention is the radical of a monocyclic or bicyclic (C3-C9)-heteroaromatic which contains one or two N atoms and/or an S or an O atom in the ring system. For the term xe2x80x9cheteroaromaticxe2x80x9d, see Garrat, Vollhardt, Aromatizitxc3xa4t [Aromaticity], Stuttgart 1973, pages 131-153. Examples of suitable heteroaryl radicals are the radicals of thiophene, furan, benzo[b]thiophene, benzofuran, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indole, quinoline, isoquinoline, oxazole, isoxazole, thiazole, isothiazole, isobenzofuran, indolizine, isoindole, indazole, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline and furazan.
As indicated above, aryl, alkyl, heteroaryl and radicals derived therefrom can be monosubstituted or, if chemically possible, alternatively polysubstituted.
Suitable polymeric solid supports are, for example, crosslinked polystyrenes (e.g. aminomethylpolystyrene (AMPS) or Tentagel.
If not stated otherwise, chiral centers can be present in the R or in the S configuration. The invention relates both to the optically pure compounds and to mixtures of stereoisomers such as mixtures of enantiomers and mixtures of diastereomers.
Suitable salts are, in particular, alkali metal and alkaline earth metal salts, salts with physiologically tolerable amines and salts with inorganic or organic acids such as, for example, HCl, HBr, H2SO4, maleic acid, fumaric acid.
The abovementioned compounds of the formulae I and II and IIa, IIb or IIc respectively are derivatives of tetrahydropyran which can be synthesized rapidly and in automated form in good yields and high purities on a solid phase with the aid of the combinatorial method described here.
Compounds of the formula I can be prepared, for example, with the aid of intermediates of the formula II which have an orthogonal protective group pattern with one or preferably a number of protective groups from the following different classes:
base-labile protective groups, such as the acetate or benzoyl group;
acid-labile protective groups, such as acetal- or ketal-like protective groups such as the ethoxyethyl group;
fluoride-labile protective groups, such as the tert-butyldimethylsilyl or tert-butyldiphenylsilyl group;
a protective group which can be removed by transition metal catalysis, such as the allyl group;
sulfur-containing protective groups, such as in the linker system.
The invention thus relates to a process for the preparation of compounds of the formula I 
and their physiologically tolerable salts, in which:
R1, R2, R3, R4, R5 independently of one another are
1. hydrogen;
2. (C1-C12)-alkyl;
3. (C2-C8)-alkenyl;
4. (C2-C8)-alkynyl;
5. (C1-C6)-alkylene-(C3-C10)-cycloalkyl;
6. (C0-C6)-alkylene-(C6-C12)-aryl; preferably phenyl or benzyl;
7. (C1-C6)-alkoxy;
8. (C0-C6)-alkylene-COxe2x80x94R8;
9. (C1-C6)-alkylene-(C1-C9)-heteroaryl;
10. carbamoyl;
11. xe2x80x94C(O)NR6R7;
12. xe2x80x94C(O)OR6;
13. a radical defined as in 2.-12., which is mono-, di- or polysubstituted in the alkyl moiety and/or aryl or heteroaryl moiety by a radical from the group consisting of (C1-C6)-alkyl, NO2, CN, halogen, CF3 or (C1-C6)-alkoxy;
14. a radical defined as in 6. and 9., which is substituted in the aryl or heteroaryl moiety by one, two or more halogen atoms; or
R2 and R3 together or R3 and R4 together or R4 and R5 together are (C1-C3)-alkylene which can be substituted by 1 or 2 (C1-C3)-alkyl radicals or optionally substituted (C6-C12)-aryl radicals;
R6 and R7 independently of one another are:
1. hydrogen;
2. (C1-C12)-alkyl;
3. (C2-C8)-alkenyl;
4. (C2-C8)-alkynyl;
5. (C1-C6)-alkylene-(C3-C10)-cycloalkyl;
6. (C1-C6)-alkylene-(C6-C12)-aryl; preferably benzyl;
7. (C2-C6)-alkyloxy;
8. (C0-C6)-alkylene-COxe2x80x94R8;
9. (C1 -C6)-alkylene-(C1-C9)-heteroaryl;
10. (C0-C6)-alkylene-(C1-C6)-alkoxy;
11. (C3-C10)-cycloalkyl;
12. (C6-C12)-aryl, preferably phenyl;
R8 is hydrogen, (C1-C6)-alkyl, (C6-C12)-aryl or OR12;
R12 is hydrogen, (C1-C6)-alkyl or (C6-C12)-aryl; and
X is N or O; by
a) introduction of a suitable, preferably sulfur-containing linker on the anomeric center of an unprotected, partially orthogonally protected or completely orthogonally protected monosaccharide derivative of the formula 
xe2x80x83in which R in each case independently of one another is hydrogen or a protective group customary in sugar chemistry; and X is O or N, preferably O;
b) reaction of a compound linker-bonded in this way, to give compounds of the formula II 
xe2x80x83in which R1 is a linker group which can be linked via a covalent bond to a support functionalized by a heteroatom, for example N, O or Cl, and
R2, R3, R4, R5 independently of one another are a protective group customary in sugar chemistry, Y is O or S, preferably S and X is O or N, preferably O, by successive or simultaneous introduction of protective groups onto the functional groups xe2x80x94OR or xe2x80x94Xxe2x80x94R, the protective groups belonging to identical or different, preferably different, orthogonal protective group classes;
c) linkage of the monosaccharide derivative of the formula II protected in this way to a polymeric solid support via the linker;
d) selective deprotection of the functional group to be derivatized on the polymeric solid support;
e) derivatization of the deprotected functional groups on the polymeric solid support, where the deprotection and subsequent derivatization of the different functional groups can be carried out selectively and also a number of equally protected functional groups can be deprotected and derivatized simultaneously;
f) removal of the derivatives bonded to the polymeric solid support, for example by activation of the sulfur on the anomeric center by bromine, and subsequent conversion of the compound activated in this way into a compound of the formula I derivatized on the anomeric center.
For the synthesis of the selectively protected monosaccharide derivatives according to formula II or IIa, IIb or IIc respectively on a solid phase, linkage to the anomeric center via a thioglycoside or an O-glycoside, in particular via a thioglycoside, is suitable. In this case, the different monosaccharides such as, for example, glucose, galactose or mannose, differ only slightly in the design of the protective groups and the sequence of their introduction. The differences in the reactivity of the functional groups and the differences associated therewith in the sequence of introduction of the different protective groups is a known problem in carbohydrate chemistry.
The synthesis strategy for the preparation of compounds of the formulae I and II is illustrated in scheme 1 by the example of the glucose derivative and is also transferable to other monosaccharides such as, for example, galactose (see scheme 3) or mannose (see scheme 4) with the abovementioned slight variations.
Compounds of the formula I can also be prepared by carrying out the linkage of selectively protected compounds of the formula II to a polymeric solid support by means of another OH position, for example the 6 OH position, such as shown in scheme 6 by the example of galactose. The linkage of the linker via the 6-OH position is possible, for example, according to the synthesis route shown in scheme 5.
Bonding of a Glycosaccharide to the Support Material (Scheme 1)
Reaction of the known 3-0-allyl-protected glucose P-acetate 3 (K. Takeo et al., Carbohydrate Research 133, 1984, 275) with the succinimide linker 2 prepared from 1 leads to compound 4. The xcex2-configured thioglycoside 7 can be prepared analogously from the N-acylated cysteamine 6 by reaction with 3 under BF3 catalysis. The deacetylation of 4 and 7 affords 8 homogeneously. Silylation on the C-6 hydroxyl group 9 and introduction of the ethoxyethyl protective group affords 10. Hydrolysis of the imide structure in 10 and coupling of the resulting acid 11 to a suitable support such as, for example, aminomethylpolystyrene affords the resin 12 loaded with the protected monosaccharide.
Reaction on the Solid Phase (see Scheme 2)
The removal and reaction of the protected monosaccharide 12 on the solid phase to give compounds of the formula I (13) is shown by way of example in scheme 2. The C-2 hydroxyl function is deacetylated by reaction with hydrazine, the hydroxyl function can then be activated by reaction with potassium tert-butoxide or phosphazene as a base (R. Schwesinger, H. Schlemper, Angew. Chem. 99, 1987, 1212-1214). The activated derivative is trapped by means of electrophiles. An analogous reaction can be carried out in the case of a C-2 amino function. The protective group used here is, for example, the Fmoc group, which can be removed by piperidine.
The removal of the allyl ether on the C-3 hydroxyl group is carried out under zirconocene catalysis (E. Negishi, Tetrahedron Lett. 1986, 27, 2829-2832; E. Negishi, Synthesis, 1988, 1-19). The functionalization is carried out analogously to the manner described above. By this means, the use of strong acids, such as would be necessary in other removal methods familiar to the person skilled in the art, can be avoided and the orthogonality to the other protective groups is guaranteed.
Alternatively to the base-catalyzed functionalization, the allyl ether protective group can be converted into a propyl group by reduction with diimine (see Hxc3xcning, H. R. Mxc3xcller, W. Thier, Angew. Chem. 1965, 77, 368-377). The C4 hydroxyl function can be removed by transacetalization in an analogous manner to that used in the case of THP acetals (cf. E. J. Corey, H. Niwa, J. Knolle, J. Am. Chem. Soc. 1978, 100, 1942-1943). The functionalization is carried out as described above. The C-6 hydroxyl function is desilylated by reaction with fluoride ions; the reaction with electrophiles is carried out analogously to C-2.
The individual steps can be carried out in a different sequence on account of the compatibilities.
After completion of the functionalization of the various groups, the anomeric position is activated by reaction of the polymerically bonded selectively protected monosaccharide with bromine/di-tert-butylpyridine. The 1-bromo derivative is converted into a derivative functionalized on the anomeric center by reaction with alcohol.
Synthesis Sequence for the Preparation of Galactose Derivatives (see Scheme 3)
Starting from galactose pentaacetate 14, the thioglycoside 16 is prepared by boron fluoride-catalyzed reaction with 15. Reaction with sodium methoxide affords 17 with deacetylation. The selective silylation of 17 takes place on the C-6 hydroxyl function. The silyl ether 18 is converted into the isopropylidene-protected derivative 19 using dimethoxypropane. Acetylation by reaction with acetic anhydride affords 20. After removal of the isopropylidene protective group, the dihydroxy compound is converted into the allyl ether derivative protected on C-4 using dibutyltin oxide and allyl bromide. Introduction of the ethoxyethyl protective group affords 21. The ester is hydrolyzed with lithium hydroxide analogously to glucose.
Synthesis Sequence for the Preparation of Mannose Derivatives (see Scheme 4)
Mannose pentaacetate 22 is reacted with thiol 6 with boron trifluoride catalysis to give the thiomannoside 23. Removal of the acetate protective groups by sodium methoxide affords 24. Reaction with dimethoxybenzaldehyde affords the acetal 25. Reaction with dibutyltin oxide and allyl bromide leads to the 3-O-allyl ester. By acetylation with acetic anhydride, 26 is prepared. The removal of the ketal and subsequent selective silylation on C-6 affords a silyl ether. Introduction of the ethoxyethyl protective group affords 27. The ester in 27 is hydrolyzed with lithium hydroxide analogously to glucose. 
Bonding to a Polymeric Solid Support via the 6-OH Position (Scheme 5) 
For anchorage via the primary hydroxyl group, a thioglycoside 28, for example, can be used as a starting material. The removal of the TBDPS group is carried out with tetrabutylammonium fluoride in THF. For the introduction of the linker, for example, etherification according to Mitsunobu is possible. The protection of the 4-hydroxyl group in 30 is carried out with slight warming with SEM-CI and Hujnig""s base in dichloromethane.
Synthesis Sequence for the Preparation of Galactose Derivatives Which are Bonded to a Polymeric Solid Support via the 1-OH Group (Scheme 6)
After binding of the glycoside to the solid polymeric support, the 1-OH group and the 2-OH group are first to be functionalized in the desired sequence, then the SEM group and the allyl ether are removed successively and the 3- and 4-positions are derivatized. The detachment of the solid polymer, for example using cerium ammonium nitrate (CAN), and the functionalization of the 6-OH function are then carried out. 
Coupling of a methyl galactosylmercaptobutyrate to an amino-funtionalized polymeric support via the 1-position (scheme 7) 
On account of their polyvalency and their defined spatial arrangement, the compounds of the formulae II, IIa, IIb and IIc are suitable as structural units for biological mimetics, for example peptide mimetics, and are a useful aid for the preparation and/or discovery of new lead structures and the identification of potential active compounds.
The compounds of the formula I prepared with the aid of the compounds of the formula II, IIa, IIb or IIc have potential diagnostic and/or pharmacological action in various forms of disorder. Autoimmune diseases and carcinomatous disorders, for example, may be mentioned.
On account of their potentially valuable pharmacological properties, the compounds according to the present invention and their physiologically tolerable salts are very highly suitable for use as therapeutics in mammals, in particular man.
The present invention therefore furthermore relates to a pharmaceutical comprising one or more compounds of the formula I and/or its pharmacologically tolerable salts, and their use for the production of a pharmaceutical for the therapy or prophylaxis of autoimmune diseases, for example rheumatism or carcinomatous disorders.
The pharmaceuticals are particularly suitable for the treatment of acute and chronic inflammation, which can be characterized pathophysiologically by a disorder of the cell circulation, for example of lymphocytes, monocytes and neutrophilic granulocytes. These include autoimmune disorders such as acute polyarthritis, rheumatoid arthritis and insulin-dependent diabetes (diabetes mellitus IDDM), acute and chronic transplant rejection, shock lung (ARDS, adult respiratory distress syndrome), inflammatory and allergic skin disorders such as, for example, psoriasis and contact eczema, cardiovascular disorders such as myocardial infarct, reperfusion injuries after thrombolysis, angioplasty or by-pass operations, septic shock and systemic shock. A further potential indication is the treatment of metastasizing tumors. Moreover, these pharmaceuticals, which are stable in the acid medium of the stomach, can be employed for the antiadhesive therapy of Helicobacter pylori and related microorganisms, if appropriate also in combination with antibiotics. Therapy of the cerebral form of malaria is furthermore conceivable with the aid of these pharmaceuticals.
Further potential application possibilities of the pharmaceuticals are in the treatment of metabolic disorders, such as diabetes and arteriosclerosis, of disorders of the cardiovascular and the central nervous system and of disorders of bone metabolism, and in their use as an antiinfective or as a pharmaceutical having immunomodulating properties.
Pharmaceuticals which contain a compound of the formula I can be administered here orally, parenterally, intravenously, rectally or by inhalation, the preferred administration being dependent on the particular course of the disorder. The compounds I can be administered here on their own or together with pharmaceutical excipients, to be specific both in veterinary and in human medicine.
The person skilled in the art is familiar on the basis of his expert knowledge with the auxiliaries which are suitable for the desired pharmaceutical formulation. In addition to solvents, gel-forming agents, suppository bases, tablet auxiliaries, and other active compound carriers, it is possible to use, for example, antioxidants, dispersants, emulsifiers, antifoams, flavor corrigents, preservatives, solubilizers or colorants.
For an oral administration form, the active compounds are mixed with the additives suitable therefor, such as vehicles, stabilizers or inert diluents, and brought into the suitable administration forms, such as tablets, coated tablets, hard gelatin capsules, aqueous, alcoholic or oily solutions, by the customary methods. Inert carriers which can be used are, for example, gum arabic, magnesia, magnesium carbonate, potassium phosphate, lactose, glucose or starch, in particular corn starch. In this case, preparation can take place both as dry and as moist granules. Suitable oily vehicles or solvents are, for example, vegetable or animal oils, such as sunflower oil or cod liver oil.
For subcutaneous or intravenous administration, the active compounds, if desired with the substances customary therefor, such as solubilizers, emulsifiers or further excipients, are brought into solution, suspension or emulsion. Suitable solvents are, for example: water, physiological saline solution or alcohols, e.g. ethanol, propanol, glycerol, in addition also sugar solutions such as glucose or mannitol solutions, or alternatively a mixture of the various solvents mentioned.
Suitable pharmaceutical formulations for administration in the form of aerosols or sprays are, for example, solutions, suspensions or emulsions of the active compound of the formula I in a pharmaceutically acceptable solvent, such as, in particular, ethanol or water, or a mixture of such solvents.
If required, the formulation can also additionally contain other pharmaceutical excipients such as surfactants, emulsifiers and stabilizers and also a propellant. Such a preparation customarily contains the active compound in a concentration of approximately 0.1 to 10, in particular from approximately 0.3 to 3, % by weight.
The dose of the active compound of the formula I to be administered and the frequency of administration depend on the potency and duration of action of the compounds used; in addition also on the nature and severity of the disease to be treated and on the sex, age, weight and individual responsiveness of the mammal to be treated.
The daily dose can be administered either by single administration in the form of an individual dose unit or else of a number of small dose units and also by multiple administration of subdivided doses at specific intervals. The daily dose to be administered can moreover be dependent on the number of receptors expressed during the course of the disease. It is conceivable that in the initial stage of the disease only a few receptors are expressed on the cell surface and accordingly the daily dose to be administered is lower than in severely ill patients.
On average, the daily dose of a compound of the formula I in a patient approximately 75 kg in weight is at least 0.001 mg/kg, preferably at least 0.01 mg/kg, to at most 10 mg/kg, preferably at most 1 mg/kg, of body weight.
Leukocyte Adhesionxe2x80x94Testing of the Activity of the Compounds According to the Invention in vivo
In inflammatory processes and other conditions activating cytokines, tissue destruction by immigrating or microcirculation-blocking leukocytes plays a crucial role. The phase which is first and crucial for the further disease process is the activation of leukocytes within the blood stream, in particular in the pre- and postcapillary area. In this case, after the leukocytes have left the axial flow of the blood, a first attachment of the leukocytes to the vascular inner wall, i.e. to the vascular endothelium, occurs. All subsequent leukocyte effects, i.e. the active diffusion through the vascular wall and the subsequent orientated migration into the tissue, are secondary reactions (Harlan, J. M., Leukocyte-endothelial interaction, Blood 65, 513-525, 1985). This receptor-mediated interaction of leukocytes and endothelial cells is regarded as an initial sign of the inflammatory process. In addition to the adhesion molecules already physiologically expressed, under the action of inflammatory mediators (leukotrienes, PAF) and cytokines (TNF-alpha, interieukines) the temporally graduated, massive expression of adhesion molecules on the cells occurs. They are at present divided into three groups: 1. immunoglobulin gene superfamily, 2. integrins and 3. selectins. While the adhesion between molecules of the Ig superfamily and the protein-protein bonds proceeds, lectin-carbohydrate bonds are prominent in the cooperation between selectins (Springer, T. A., Adhesion receptors of the immune system. Nature 346, 425-434, 1990; Huges, G., Cell adhesion moleculesxe2x80x94the key to an universal panacea, Scrips Magazine 6, 30-33, 1993; Springer, T. A., Traffic signals for lymphocyte recirculation and leukocyte emigration; The multistep paradigm, Cell 76, 301-314, 1994).
The Activity of the Compounds According to the Invention in vivo can be Tested According to the Following Method:
The induced adhesion of leukocytes is quantified in the mesenterium of the rat using an intravital microscopic investigation technique (Atherton A. and Born G. V. R., Quantitative investigations of the adhesiveness of circulating polymorphonuclear leukocytes to blood vessel walls. J. Physiol. 222, 447-474, 1972; Seiffge, D. Methoden zur Untersuchung der Rezeptor-vermittelten Interaktion zwischen Leukozyten und Endothelzellen im Entzxc3xcndungsgeschehen [Methods for the investigation of receptor-mediated interaction between leukocytes and endothelial cells in the inflammation process], in: Ersatz- und Ergxc3xa4tnzungsmethoden . zu Tierversuchen in der biomedizinischen Forschung [Replacement and supplementary methods for animal experiments in biomedical research], Schxc3x6ffl, H. et al., (Ed.) Springer, 1995 (in press)). Under inhalation ether anesthesia, prolonged anesthesia is initiated by intramuscular injection of urethane (1.25 mg/kg of BW). After exposure of vessels (femoral vein for the injection of substances and carotid artery for blood pressure measurements), catheters are tied into these. The appropriate transparent tissue (mesenterium) is then exposed by standard methods known in the literature and laid out on the microscope stage and covered with a layer of paraffin oil at 37xc2x0 C. (Menger, M. D. and Lehr, H., A. Scope and perspectives of intravital microscopy-bridge over from in vitro to in vivo, Immunology Today 14, 519-522, 1993). The test substance is administered i.v. to the animal (10 mg/kg). The experimental increase in blood cell adhesion is induced by means of cytokine activation by systemic administration of lipopolysaccharide (LPS, 15 mg/kg) 15 minutes after administration of test substance (Foster S. J., McCormick L. M., Ntolosi B. A. and Campbell D., Production of TNF-alpha by LPS-stimulated murine, rat and human blood and its pharmacological modulation, Agents and Actions 38, C77-C79, 1993, 18.01.1995). The increased adhesion of leukocytes to the endothelium caused thereby is quantified directly by vital microscopy or with the aid of fluorescent dyes. All measuring processes are carried out by video camera and stored on a video recorder. Over a period of 60 minutes, the number of rolling leukocytes, (i.e. all visible rolling leukocytes, which are slower than the flowing erythrocytes) and the number of adhering leukocytes on the endothelium (residence time longer than 5 seconds) are determined every 10 minutes. After completion of the experiment, the anesthetized animals are put to sleep without excitation in a pain-free manner by systemic injection of T61. For evaluation, the results of 8 treated animals in each case are compared (in percent) with those of 8 untreated animals (control group).